THE EFFECTS OF ETANERCEPT AND CABERGOLINE ON ENDOMETRIOTIC IMPLANTS, UTERUS AND OVARIES IN RAT ENDOMETRIOSIS MODEL.

The pathophysiology of endometriosis is still unknown and treatment options remain controversial. Searches focus on angiogenesis, stem cells, immunologic and inflammatory factors. This study investigated the effects of etanercept and cabergoline on ovaries, ectopic, and eutopic endometrium in an endometriosis rat model. This randomized, placebo-controlled, blinded study included 50 rats, Co(control), Sh(Sham), Cb(cabergoline), E(etanercept), and E + Cb(etanercept + cabergoline) groups. After surgical induction of endometriosis, 2nd operation was performed for endometriotic volume and AMH level. After 15 days of treatment: AMH level, flow cytometry, implant volume, histologic scores, immunohistochemical staining of ectopic, eutopic endometrium, and ovary were evaluated at 3rd operation. All groups had significantly reduced volume, TNF-α, VEGF, and CD 146/PDGF-Rß staining of endometriotic implants comparing to the Sh group (p < 0.05).TNF-α staining of eutopic endometrium in all treatment groups was similar to Sh and Co groups (p > 0.05). E and E + Cb groups significantly decreased TNF-α staining in the ovary comparing to Sh, Co, and Cb groups (p < 0.05). All treatment groups had significantly higher AFC compared to the Sh group. CD25+ Cells' median percentage was significantly increased in the E + Cb group compared to Co, Sh, Cb, and E group. E + Cb group had a significantly higher CD5+ Cells' level than the Co group (p = 0.035). In conclusion; Etanercept and/or Cabergoline decreased volume, TNF-α, VEGF, and CD 146/PDGF-Rß staining of the ectopic endometrial implant. E and E + Cb treatment decreased TNF-α levels in the ovary. E + Cb also increased peripheral blood CD25+ & CD5+ Cell's.

Comparative Assessment of Leukocyte Infiltrate Composition in the Uterine and Vaginal Tissues in Rats with Experimental Endomyometritis Treated with Interferon-γ at Different Times of the Day.

The cell composition of leukocyte infiltrates in the endometrium, myometrium, and vaginal walls was studied in Wistar rats with modeled chronic endomyometritis after administration of IFNγ (0.1 µg/100 g body weight) in different daily regimens (10.00 or 20.00). Morning injections of this cytokine ameliorated inflammatory infiltration of the uterine wall and vagina, but increased the content of neutrophils in the endometrium. Evening cytokine injections reduced neutrophilic infiltration, enhanced mononuclear infiltration, and had no effect on plasmacytic infiltration of the uterine and vaginal walls. In the vaginal wall, both IFNγ administration schedules decreased neutrophil content. The data indicate the necessity to take into account the circadian rhythms in IFN therapy.

Potential impact of maternal vitamin D status on peripheral blood and endometrium cellular immunity in women with recurrent implantation failure.

PROBLEM: This study aims to evaluate the modulatory effects of vitamin D on peripheral blood and endometrial cellular immunity in women with recurrent implantation failure (RIF). METHOD OF STUDY: One hundred and fifty-four women with RIF were identified at a fertility center from January 2018 and March 2019. Blood and endometrium samples were collected during the mid-luteal phase before IVF treatment or pregnancy. The serum vitamin D status, NK cell cytotoxicity, Th1 cytokine production, and endometrial immune cells were detected before and after vitamin D supplementation. RESULTS: The NK cell cytotoxicity at an effector:target (E:T) ratio of 50:1 or 25:1 was significantly higher in vitamin D insufficiency group (VDI) than those in vitamin D normal group (VDN) (P < .05 each). The percentage of IFN-γ- or TNF-α-producing Th cells was significantly increased in VDI or vitamin D deficiency group (VDD) when compared with VDN (P < .05 each). The percentage of CD68+ macrophages on all endometrial cells in VDI and VDD was significantly higher than in VDN (P < .05 each), while no significant differences in the percentage of other endometrial immune cells among the three groups were observed. This dysregulation was significantly reduced with vitamin D supplementation. CONCLUSION: Our findings highlighted that vitamin D may have an important role in the regulation of not only systemic but also local immune response for optimization of maternal tolerance for implantation in women with RIF. Pre-conception optimization of vitamin D status should be considered in women with RIF.

Intrauterine infusion of autologous platelet-rich plasma in women undergoing assisted reproduction: A systematic review and meta-analysis.

Prior studies have provided conflicting results regarding the use of platelet-rich plasma (PRP) in women undergoing in-vitro fertilization (IVF) or intracytoplasmic injection (ICSI). The objective of this study was to evaluate the effect of the intrauterine infusion of PRP on the outcome of embryo transfer (ET) in women undergoing IVF/ICSI. We searched databases, including PubMed, Embase, Scopus, Web of Science, and the Cochrane Database of Clinical Trials (CENTRAL). Meta-analysis using a random-effects model was performed to calculate the pooled estimates. Seven studies involving 625 patients (311 cases and 314 controls) were included. The probability of chemical pregnancy (n = 3, risk ratio (RR): 1.79, 95 % confidence intervals (CI): 1.29, 2.50; P < 0.001, I2 = 0 %), clinical pregnancy (n = 7, RR: 1.79, 95 % CI: 1.37, 2.32; P < 0.001, I2 = 16 %), and implantation rate (n = 3, RR: 1.97, 95 % CI: 1.40, 2.79; P < 0.001, I2 = 0 %) was significantly higher in women who received PRP compared with control. There was no difference between women who received PRP compared with control group regarding miscarriage (RR: 0.72, 95 % CI: 0.27, 1.93; P = 0.51, I2 = 0 %). Following the intervention, endometrial thickness increased in women who received PRP compared to control group (SMD: 1.79, 95 % CI: 1.13, 2.44; P < 0.001, I2 = 64 %). The findings of this systematic review suggest that PRP is an alternative treatment strategy in patients with thin endometrium and recurrent implantation failure (RIF). Further prospective, large, and high quality randomized controlled trials (RCTs) are needed to identify the subpopulation that would most benefit from PRP.

Efficacy of intrauterine administration of autologous peripheral blood mononuclear cells on the pregnancy outcomes in patients with recurrent implantation failure: A systematic review and meta-analysis.

One in every nine couples suffers from implantation defects and pregnancy failures. In spite of many contributions that ART has given to infertility treatment, there are many reports of the failure of ART. Therefore, scientists suggested many complementary therapies for use besides ART to improve the quality of infertility treatments. Intrauterine PBMC-therapy is one of these complementary therapies that were used before IVF. Studies that examined PBMC treatment in women with at least three IVF/ET failure were included in this review. These studies involved RCT and quasi-experimental (non-randomized experimental) studies. A three-step search strategy was used for published and unpublished clinical trials written in English and Persian. No time limitation was set for studies. Study selection according to the inclusion criteria and methodological quality assessment and data extraction were done by two independent reviewers, which result in five studies being included (two RCTs and three quasi-experimental studies). Finally, all of these article extracted data were pooled in a statistical meta-analysis. Findings demonstrated that implantation, pregnancy and live birth rate were statistically increased and the miscarriage rate was significantly decreased in the PBMC-treated group than that non-treated group. In conclusion, based on the evidence, PBMCs can be an effective therapeutic approach in women with at least three IVF/ET failure and lacking initial inflammation that is essential for implantation.

Curcumin attenuates proangiogenic and proinflammatory factors in human eutopic endometrial stromal cells through the NF-κB signaling pathway.

Endometriosis is a chronic gynecological inflammatory disorder in which immune system dysregulation is thought to play a role in its initiation and progression. Due to altered sex steroid receptor concentrations and other signaling defects, eutopic endometriotic tissues have an attenuated response to progesterone. This progesterone-resistance contributes to lesion survival, proliferation, pain, and infertility. The current agency-approved hormonal therapies, including synthetic progestins, GnRH agonists, and danazol are often of limited efficacy and counterproductive to fertility and cause systemic side effects due to suppression of endogenous steroid hormone levels. In the current study, we examined the effects of curcumin (CUR, diferuloylmethane), which has long been used as an anti-inflammatory folk medicine in Asian countries for this condition. The basal levels of proinflammatory and proangiogenic chemokines and cytokines expression were higher in primary cultures of stromal cells derived from eutopic endometrium of endometriosis (EESC) subjects compared with normal endometrial stromal cells (NESC). The treatment of EESC and NESC with CUR significantly and dose-dependently reduced chemokine and cytokine secretion over the time course. Notably, CUR treatment significantly decreased phosphorylation of the IKKα/ß, NF-κB, STAT3, and JNK signaling pathways under these experimental conditions. Taken together, our findings suggest that CUR has therapeutic potential to abrogate aberrant activation of chemokines and cytokines, and IKKα/ß, NF-κB, STAT3, and JNK signaling pathways to reduce inflammation associated with endometriosis.

Intralipid® may represent a new hope for patients with reproductive failures and simultaneously an over-immune endometrial activation.

PROBLEM: Continuous failures to achieve a pregnancy despite effective embryo transfers is extremely distressing for couples. In consequence, many adjuvant therapies to IVF have been proposed to achieve an "ideal" immune environment. We here focus on Intralipid® therapy (IL) reported to have immunosuppressive properties on NK cells. METHOD OF STUDY: 94 patients exhibited an immune profile of endometrial over-immune activation and an history of repeated implantation failures despite multiple embryos transfers (RIF). They received a slow perfusion of Intralipid®. We here report the live birth rate following the procedure at the next embryo transfer. To get new insight on its mechanism of action, a second immune profiling had been performed under Intralipid® before the embryo transfer. RESULTS: The live birth rate of the RIF cohort treated with Intralipid® reached 54% (51/94) at the next embryo transfer. In patients successfully pregnant under Intralipid® who benefitted of a test of sensibility before the embryo transfer, we observed a significant decrease of the three biomarkers used to diagnose the over-immune endometrial activation (CD56 cells; IL-18/TWEAK, IL-14/FN-14). CONCLUSIONS: Double blind placebo versus Intralipid® studies should be conducted. Intralipid® may be an option to explore in RIF patients who exhibit an over-immune activation of uNK cells.

Vitamin D Regulates Maternal T-Helper Cytokine Production in Infertile Women.

Vitamin D (VD) deficiency is associated with reproductive failure. However, the relationship between VD and maternal immunity remains unclear. We investigated the clinical efficacy of VD in maternal T-helper (Th) cytokines in 276 infertile women and examined for Th1 and Th2 cells based on the deficient, insufficient, and sufficient serum 25-hydroxyvitamin D3 (25[OH]VD) levels (<12, 12⁻30, and >30 ng/mL, respectively). Most infertile women had a low-level of VD (87.3%). Immunological tests of pre-/post-VD supplementation were performed in patients who were deficient and insufficient in VD. Of 23 patients, 11 (47.8%) exhibited sufficient VD levels after supplementation. Th1/Th2 cell ratio in patients with insufficient VD was significantly decreased after supplementation (p = 0.004). After supplementation, serum 25(OH)VD levels of the patients: 11 in the sufficient group showed significant decreases in Th1 cell level and Th1/Th2 cell ratio (p = 0.032 and 0.010, respectively), whereas no significant differences in Th1/Th2 cell ratio were recognized in the insufficient group. Furthermore, mid-luteal endometrial biopsies (n = 18) were processed for primary cultures and measured interferon [IFN]-γ and interleukin [IL]-4 in condition media. Decidualizing cultures with 1,25-dihydroxvitamin D3 (1,25[OH]2VD) decreased IFN-γ. Sufficient VD supplementation in women with insufficient VD may optimize maternal T-helper cytokines during pregnancy via rebalancing the Th1/Th2 cell ratio.

Effect of Xianziyizhen Recipe Capsule on PGI2-PPARδ Signaling Pathway in Embryo Implantation Dysfunction Mice.

PROBLEM: We investigated the effect of Xianziyizhen recipe capsule (XRC), a kidney-tonifying herb, on the PGI2-PPARδ signaling pathway at the maternal-fetal interface in embryo implantation dysfunction (EID) mice. METHOD OF STUDY: Intragastric administration of Progynova (estradiol) or XRC was performed in EID mouse model, following experimental induction of kidney deficiency by co-treatment with chemotherapy drug hydroxyurea and antiprogesterone mifepristone. The PPARδ and IL-11 mRNA expression in endometrium were detected by real-time relative reverse transcription-polymerase chain reaction (RT-PCR). Further, the protein expression of COX-2, PGI2, MMP-9, and TIMP-3 was detected in endometrial glandular epithelium and in stromal cells by immunohistochemical (IHC) assay. RESULTS: The results showed that hydroxyurea and mifepristone-induced EID were associated with significantly lower PPARδ and IL-11 mRNA levels in endometrium and reduced COX-2, PGI2, MMP-9, and TIMP-3 levels in endometrial glandular epithelium, compared with normal controls. However, XRC and Progynova treatment reversed these effects, leading to significant increases in PPARδ and IL-11 mRNA expression, and COX-2, PGI2, MMP-9 and TIMP-3 protein levels, when compared with the levels observed in EID mice. CONCLUSION: These results strongly suggested that XRC is beneficial in EID treatment and that XRC may mediate its effects through regulation of the PGI2-PPARδ signaling pathway.

A randomised controlled trial of a preconceptional dietary intervention in women undergoing IVF treatment (PREPARE trial).

BACKGROUND: In vitro fertilisation (IVF) treatment provides an opportunity to study early developmental responses to periconceptional dietary interventions. Retrospective studies have suggested links between preconception diet and fertility, and more recently, a "Mediterranean" diet has been reported to increase pregnancy rates by up to 40%. In addition, a prospective study examining increased intake of omega-3 polyunsaturated fats demonstrated a quickened rate of embryo development after IVF. However, up to now, few prospective randomised controlled trials have investigated the impact of periconceptional dietary interventions on fertility outcomes. METHODS AND DESIGN: The study is a randomised controlled trial of a dietary intervention consisting of olive oil for cooking, an olive oil based spread, and a daily supplement drink enriched with Vitamin D (10 microgram daily) and marine omega-3 fatty acids (2 g daily) for 6 weeks preconception versus a control diet of sunflower seed oil for cooking, a sunflower oil based spread, and a daily supplement drink without added Vitamin D or marine omega-3 fatty acids. Couples undergoing IVF will be randomised to either the intervention or control group (55 in each arm). The primary endpoint is embryo developmental competency in vitro, measured by validated morphokinetic markers. Secondary outcomes will include the effect of the dietary intervention on the nutritional content of the intrauterine environment. DISCUSSION: This approach will enable rigorous examination of the impact of the dietary intervention on early embryo development, together with the influence of the peri-implantation intra-uterine nutritional environment. TRIAL REGISTRATION: ISRCTN50956936.

[Research of therapeutical effect and immunologic mechanism of Jiawei Foshou San on model rats of endometriosis].

OBJECTIVE: To explore the effect and mechanism of Jiawei Foshou San on the accretion ectopic endometrium of rats. METHOD: Endometriosis model was established by surgical implant of endometrial tissue which belongs to its body in rats. Jiawei Foshou San was administrated to the model rats. Twenty-eight days later, the length of ectopic endometrium was measured by vernier caliper, the spleen was weighed by electronic balance, the content of tumor necrosis factor (TNF)-alpha in blood serum and peritoneal fluid was detected by ELISA test, the expression of interleukin (IL)-8 in ectopic endometrium was detected by immunohistochemical method, the expression of NF-kappaB p65 protein and inhibitory KBalpha (IkappaBalpha) protein in ectopic endometrium were analyzed by western blot. RESULT: Jiawei Foshou San 0.045, 0.09, 0.18 g x kg(-1) group reduced the volume of ectopic endometrium. Jiawei Foshou San 0.18 g x kg(-1) group raised the spleen exponent of model rats. Jiawei Foshou San 0.09, 0.18 g x kg(-1) group decreased the content of TNF-alpha in blood serum and peritoneal fluid, and the content of IL-8 in ectopic endometrium was also decreased. Jiawei Foshou San can decrease the expression of NF-KB p65 and increase the expression of IkappaBalpha in ectopic endometrium. CONCLUSION: Jiawei Foshou San can inhibit the accretion of endometriosis implants of rats, and its mechanism might be associated with improving the environment of body.

Local immune regulatory effects of Bangdeyun on the endometrium of mice with embryo implantation dysfunction during the implantation time.

This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB (NF-kappaB), interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) in the endometrium of mice with embryo implantation dysfunction (EID) during the implantation time (namely on pregnancy day 5, 6, 7 and 8) and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-kappaB was detected by immunohistochemistry and Western blotting. IFN-gamma and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that in the normal group, NF-kappaB and IFN-gamma were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-kappaB and IFN-gamma were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group (P<0.01 for all). In the Bangdeyun-treated group, little amount of NF-kappaB and IFN-gamma was expressed and IL-10 expression was strong, much the way they were expressed in the normal group (P>0.05). The expressions of NF-kappaB and IFN-gamma were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 (P<0.01 for all), while the expression of IL-10 was much higher than in the model group during the whole implantation time (P<0.01). It was suggested Bangderun may favor a shift from Th1- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.

Local immune regulatory effects of Bangdeyun on the endometrium of mice with embryo implantation dysfunction during the implantation time / 华中科技大学学报（医学）（英德文版）

This study examined the effects of Bangdeyun on the expressions of nuclear factor-kappaB (NF-kappaB), interferon-gamma (IFN-gamma) and interleukin-10 (IL-10) in the endometrium of mice with embryo implantation dysfunction (EID) during the implantation time (namely on pregnancy day 5, 6, 7 and 8) and explored the local immune regulatory effects of Bangdeyun. The gestational mice were randomly divided into normal group, model group and Bangdeyun-treated group. EID models of mice were established by using indomethacin. The endometrial expression of NF-kappaB was detected by immunohistochemistry and Western blotting. IFN-gamma and IL-10 were measured by enzyme-linked immunosorbent assay (ELISA). The results showed that in the normal group, NF-kappaB and IFN-gamma were weakly expressed and IL-10 was strongly expressed in the endometrium during the whole implantation period. In the model group, the expressions of NF-kappaB and IFN-gamma were increased on pregnancy day 5, 6 and 7, and IL-10 expression decreased during the whole implantation time when compared with those in the normal group (P0.05). The expressions of NF-kappaB and IFN-gamma were much lower in the Bangdeyun-treated group than those in the model group on pregnancy day 5, 6 and 7 (P<0.01 for all), while the expression of IL-10 was much higher than in the model group during the whole implantation time (P<0.01). It was suggested Bangderun may favor a shift from Th1- to Th2-type immune response, therefore inhibiting the maternal immune rejection, inducing the immune tolerance and improving the fetal implantation.

[Observation on effect of combined therapy of neiyi pill and neiyi enema on endometriosis].

OBJECTIVE: To observe the effect of combined therapy of Neiyi pill (NP) and Neiyi enema (NE) on endometriosis and its effect on serum levels of endometrial antibody (EMAB) and carcinoembryonic antigen 125 (CA125). METHODS: Fifty-eight cases with endometriosis were divided into 3 groups randomly, group A (n = 16) treated by NP, group B (n = 24) treated by NP and NE, and group C (n = 18) treated by danazol, all for 3 menstrual cycle with single blind method. The effect was observed and the serum levels of EMAB and CA125 were detected before and after treatment. RESULTS: Comparison of the efficacy between the 3 groups showed: there was no remarkable difference between group A and B (P >0.05), both of group A and group B were superior to that of group C (P<0.05). The levels of EMAB and CA125 had no significant changes in all the three groups after treatment. CONCLUSION: Combined therapy of NP and NE could improve the curative effect on endometriosis, and without obvious effects on serum levels of EMAB and CA125.

[Study on mechanism of Quyujiedu method in treating endometriosis of stasis-toxic syndrome type].

OBJECTIVE: To observe the mechanism of Quyujiedu (QYJD, a method for removing stasis and detoxicating) in treating endometriosis of stasis-toxic syndrome type. METHODS: Clinical study: Sixty patients were randomly divided into two groups, the treated group treated with QYJD and the control group treated with Danazol. The changes of clinical symptoms, physical signs, pregnancy rate, as well as levels of interleukin-1 (IL-1), interleukin-6 (IL-6) and endometrial antibody (EMAb) in them before and after treatment were observed. Experimental study: Sixty rats were established into endometriosis model, and randomly divided into the QYJD treated group, the Danazol treated group for positive control and the untreated group for negative control to observe the expression of Bax. RESULTS: Clinical study showed that in the treated group and the control group, after treatment, dysmenorrhea score were 2.40 +/- 1.85 and 3.47 +/- 2.03, the menoxenia score 1.67 +/- 2.04 and 3.73 +/- 1.72 (P < 0.05), and the pregnancy rate 63.2% and 23.5% respectively, the difference between them was significant (all P < 0.05). Both of the two treatments can rectify immunity, reduce levels of IL-1 and IL-6, and lower the positive rate of EMAb. Experimental study showed that QYJD could improve the expression of Bax in ectopic endometrium, but could not affect the on-side expression of Bax in endometrium and muscle tissue. CONCLUSION: QYJD is effective in improving the stasis-toxic syndrome of endometriosis, its mechanism related with the regulation of the immune condition and the promotion of cell apoptosis in ectopic endometrium, indicating that QYJD is an effective remedy for endometriosis of stasis-toxic syndrome type.

Clinical observation on treatment of 2,062 cases of immune infertility with integration of traditional Chinese medicine and western medicine.

To study the therapeutic effect of integrated traditional Chinese medicine and western medicine on female immune infertility. 3,496 women suffering from primary or secondary infertility had their ASAb, EMAb, AOAb and ACAb level tested, with the positive rate of 23.11%, 34.95%, 20.77% and 30.41% respectively. 2,062 positive cases were periodically treated with the Chinese drug Xiaokangwan plus dexamethasone, vitamin E and vitamin C for 2 periods as a course of treatment. At the end of a treatment course, the rate for the antibodies to turn negative reached over 85% and the average pregnant rate reached 36.66%. The treatment of immune infertility with the integrated approach can reduce or eliminate the influence of antibodies in the serum of patients on various links of pregnancy, thus reaching the goal of curing infertility.

Increase in peripheral blood mononuclear cell (PBMC)- and CD56+ cell-mediated killing of endometrial stromal cells by mycobacteria; a possible role in endometriosis immunotherapy?

BACKGROUND: Immunological therapies have shown promising results in the treatment of endometriosis. Mycobacteria are one of the most common immune therapies used in other diseases. We have assessed the effects of mycobacteria in altering the ability of peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells to kill endometrial stromal cells using an in vitro model. This may have implications in the immunotherapy of endometriosis. METHODS: Primary cultures of endometrial stromal cells were grown from female patients and PBMCs were extracted from healthy female volunteers. Effector cells (PBMCs or NK cells) were exposed to varying concentrations of mycobacteria before their ability to kill cultured endometrial cells was tested using a 51Cr-release assay. RESULTS: Treatment of effector cells with the Connaught Substrain Bacillus of Calmette and Guérin (BCG) led to increased killing of target cells by PBMCs and NK cells. The optimal concentration for treatment of effector cells with Connaught BCG was approximately 0.1 multiplicities of infection (m.o.i.). There was a trend towards increased killing after treatment with Pasteur BCG. CD56+ (NK) cells treated with BCG at 0.1 m.o.i. showed increased killing of target cells compared with untreated effector cells. CONCLUSIONS: Endometrial stromal cells are susceptible to killer cells activated by mycobacteria. This in vitro work suggests a possible role for mycobacteria in the immunotherapy of endometriosis.

Anti-endometrial IgM autoantibodies in endometriotic patients: a preliminary study.

Japanese herbal medicines (Kampo medicines) are usually the third most popular choice among medicines for treatment of endometriosis in Japan. This traditional therapy is used to improve various signs and symptoms of endometriosis without decreasing serum estradiol levels or causing menstrual disorders. We used flow cytometry to examine and compare the effects of the Kampo therapy and danazol on anti-endometrial humoral immunity. Autoantibodies against endometrial epithelial cell lines and endometrial stromal cells were detectable in all the examined sera of men and women irrespective of the presence of endometriosis. Moreover, no significant increase in anti-endometrial antibodies was found in endometriotic patients. Anti-endometrial antibodies included Ig-gamma chain, Ig-mu chain, Ig-kappa chain, and Ig-lambda chain indicating polyclonal B cell activation in the endometriotic patients. Absorption tests of nonspecific antibodies with cervical cancer cells or ovarian cancer cells revealed that endometriotic patients had higher levels of endometrium-specific autoantibodies than did non-endometriotic healthy women. IgM fractions from endometriotic patients and healthy women differed in their effect on growth of endometrial adenocarcinoma cells. Therapy with the herbal compounds Keishi-bukuryo-gan but not danazol therapy, gradually decreased the tissue-specific anti-endometrial IgM antibody levels. These results indicate that tissue-specific anti-endometrial IgM may be a useful therapeutic marker for endometriotic patients treated with Keishi-bukuryo-gan and that endometrial tissue-specific immune disorders play specific roles in the pathogenesis or development of endometriosis.

The transcription of corticotropin-releasing hormone in human endometrial cells is regulated by cytokines.

Corticotropin-releasing hormone (CRH), a hypothalamic neuropeptide, is also produced in the human endometrium where it participates in local inflammatory phenomena associated with the decidualization of endometrial stroma and the implantation of the fertilized egg. The inflammatory cytokines interleukin 1 (IL-1), IL-6 and leukemia inhibitory factor (LIF) appear to be the dominant local regulators of these intrauterine inflammatory processes. In the present study we have examined the direct interactions between cytokines and CRH in the endometrium. For this purpose we have measured the effects of IL-1, IL-6 and LIF on the activity of CRH promoter inserted in human endometrial cells in culture. Homologous transient transfection experiments were conducted employing a 0.9-kb fragment of the 5' flanking region of the human CRH gene coupled to the luciferase reporter gene, using Ishikawa human endometrial cells. We have found that IL-1beta increased the activity of CRH gene promoter, in a time- and dose-dependent manner. This effect was antagonized by the IL-1 receptor antagonist IL-1ra and blocked completely by the cyclo-oxygenase inhibitor indomethacin. Similarly, IL-6 increased the activity of CRH promoter in a dose-dependent fashion, an effect partially reversed by indomethacin. LIF did not have any apparent effect. In conclusion, our data suggest that IL-1 and IL-6 exert a strong stimulatory effect on the expression of endometrial CRH. This effect is most probably mediated via prostaglandins. Based on these data we hypothesize that in the human endometrium interleukins, prostaglandins and CRH form a local network regulating the inflammatory phenomena taking place within the uterine cavity.

Effects of high dose progestogens on white cells and necrosis in human endometrium.

Endometrium was collected from 20 high-dose progestogen-treated patients and examined for leukocyte populations by immunohistochemistry and phloxine-tartrazine staining. A labelled streptavidin-biotin-alkaline phosphatase technique was used with antibodies against leukocyte common antigen (LCA), T cells (CD3), neutrophils (NP57), macrophages] (CD68, KP1) and B cells (CD20). The numbers of LCA (1070 +/- 117/mm2), CD3 (459 +/- 60/mm2), CD68 (129 +/- 21/mm2) positive cells and endometrial granulated lymphocytes (EGL) (236 +/- 41/mm2) were significantly higher than those in the control group (P < 0.001). Of these, EGL increased most (6.7 times). NP57 positive (NP57+) neutrophils were present in five out of 20 progestogen-treated samples and NP57 negative (NP57-) neutrophils in another six out of 20; while a neutrophil was only identified in one control tissue (P = 0.002). Three progestogen-exposed endometrial samples had either focal or extensive necrosis, and many NP57+ and NP57- neutrophils were present in the necrotic areas. EGL, neutrophils and macrophages are known to release a number of cytolytic and cell toxic molecules which may play a role in the initiation or acceleration of progestational endometrial necrosis.